Institut de Recherche sur la Biologie de l'Insecte (IRBI), UMR7261 CNRS - Université de Tours, Tours, France.
Institut de Systématique, Évolution, Biodiversité (ISYEB), UMR7205 Muséum National d'Histoire Naturelle, CNRS, Sorbonne Université, EPHE, Université des Antilles, Paris, France.
PeerJ. 2023 Oct 10;11:e16022. doi: 10.7717/peerj.16022. eCollection 2023.
Broad-scale monitoring of arthropods is often carried out with passive traps ( Malaise traps) that can collect thousands of specimens per sample. The identification of individual specimens requires time and taxonomic expertise, limiting the geographical and temporal scale of research and monitoring studies. DNA metabarcoding of bulk-sample homogenates has been found to be faster, efficient and reliable, but the destruction of samples prevents validation of species occurrences and relative abundances. Non-destructive metabarcoding of DNA extracted from collection medium has been applied in a limited number of studies, but further tests of efficiency are required with different trap types and collection media to assess the consistency of the method.
We quantified the detection rate of arthropod species when applying non-destructive DNA metabarcoding with a short (127-bp) fragment of mitochondrial COI on two combinations of passive traps and collection media: (1) water with monopropylene glycol (HO-MPG) used in window-flight traps (WFT, 53 in total); (2) ethanol with monopropylene glycol (EtOH-MPG) used in Malaise traps (MT, 27 in total). We then compared our results with those obtained for the same samples using morphological identification (for WFTs) or destructive metabarcoding of bulk homogenate (for MTs). This comparison was applied as part of a larger study of arthropod species richness in silver fir ( Mill., 1759) stands across a range of climate-induced tree dieback levels and forest management strategies.
Of the 53 HO-MPG samples from WFTs, 16 produced no metabarcoding results, while the remaining 37 samples yielded 77 arthropod MOTUs in total, of which none matched any of the 343 beetle species morphologically identified from the same traps. Metabarcoding of 26 EtOH-MPG samples from MTs detected more arthropod MOTUs (233) than destructive metabarcoding of homogenate (146 MOTUs, 8 orders), of which 71 were shared MOTUs, though MOTU richness per trap was similar between treatments. While we acknowledge the failure of metabarcoding from WFT-derived collection medium (HO-MPG), the treatment of EtOH-based Malaise trapping medium remains promising. We conclude however that DNA metabarcoding from collection medium still requires further methodological developments and cannot replace homogenate metabarcoding as an approach for arthropod monitoring. It can be used nonetheless as a complementary treatment when enhancing the detection of soft-bodied arthropods like spiders and Diptera.
通常使用被动陷阱(巴氏诱捕器)对节肢动物进行大规模监测,每个样本可收集数千个标本。个体标本的鉴定需要时间和分类学专业知识,限制了研究和监测研究的地理和时间范围。对批量样本匀浆进行 DNA metabarcoding 已被证明更快、更有效和更可靠,但样本的破坏阻止了对物种出现和相对丰度的验证。从收集介质中提取的 DNA 的非破坏性 metabarcoding 已在有限数量的研究中得到应用,但需要使用不同的陷阱类型和收集介质进一步测试效率,以评估该方法的一致性。
我们通过使用巴氏诱捕器(MT,共 27 个)中含有丙二醇单甲醚(EtOH-MPG)的乙醇和在窗飞诱捕器(WFT,共 53 个)中含有丙二醇单甲醚(HO-MPG)的水,对两种组合的被动陷阱和收集介质应用非破坏性短(127bp)片段线粒体 COI 的 DNA metabarcoding 时,量化了节肢动物物种的检测率。我们然后将我们的结果与使用相同样本进行形态鉴定(对于 WFT)或破坏性批量匀浆 metabarcoding(对于 MT)获得的结果进行比较。该比较是作为对银冷杉( Mill.,1759)林分中节肢动物物种丰富度的更大研究的一部分进行的,该研究涉及一系列气候诱导的树木死亡水平和森林管理策略。
在来自 WFT 的 53 个 HO-MPG 样本中,有 16 个没有产生 metabarcoding 结果,而其余 37 个样本共产生了 77 个节肢动物 MOTUs,其中没有一个与从同一诱捕器中形态鉴定的 343 种甲虫物种相匹配。来自 MT 的 26 个 EtOH-MPG 样本的 metabarcoding 检测到的节肢动物 MOTUs (233 个)多于破坏性的均质 metabarcoding (146 个 MOTUs,8 个目),其中 71 个是共享的 MOTUs,尽管两种处理方式的每个诱捕器的 MOTU 丰富度相似。虽然我们承认从 WFT 衍生的收集介质(HO-MPG)的 metabarcoding 失败,但基于乙醇的巴氏诱捕介质的处理仍然很有希望。然而,我们得出结论,DNA metabarcoding 从收集介质仍然需要进一步的方法开发,不能替代均质 metabarcoding 作为节肢动物监测的方法。但是,它可以作为补充处理方法,用于增强对蜘蛛和双翅目等软体节肢动物的检测。